The generation of inducible pluripotent stem cells (iPSCs) is a revolutionary technique allowing production of pluripotent patient-specific cell lines used for disease modeling, drug screening, and cell therapy. Integrity of nuclear DNA (nDNA) is mandatory to allow iPSCs utilization, while quality control of mitochondrial DNA (mtDNA) is rarely included in the iPSCs validation process. In this study, we performed mtDNA deep sequencing during the transition from parental fibroblasts to reprogrammed iPSC and to differentiated neuronal precursor cells (NPCs) obtained from controls and patients affected by mitochondrial disorders. At each step, mtDNA variants, including those potentially pathogenic, fluctuate between emerging and disappearing, and some having functional implications. We strongly recommend including mtDNA analysis as an unavoidable assay to obtain fully certified usable iPSCs and NPCs.

诱导型多能干细胞 (iPSC) 的产生是一项革命性的技术，允许生产用于疾病建模、药物筛选和细胞治疗的多能患者特异性细胞系。核 DNA (nDNA) 的完整性对于 iPSC 的利用是必不可少的，而线粒体 DNA (mtDNA) 的质量控制很少包含在 iPSC 验证过程中。在这项研究中，我们在从亲本成纤维细胞到重编程 iPSC 以及从对照和受线粒体疾病影响的患者获得的分化神经元前体细胞 (NPC) 的转变过程中进行了 mtDNA 深度测序。在每一步，mtDNA 变体，包括那些可能致病的变体，都会在出现和消失之间波动，有些还具有功能影响。